This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. In this project we will engineer, build, and test molecular models that will impart certain advantageous properties into enzymes of significant industrial or medical importance. For example, our B12-independent glycerol dehydratase system can be thought of as an industrial catalyst for the production of plastics from renewable resources. This is because this enzyme catalyzes the penultimate step in the production of 1,3-propanediol from glucose. We have currently filed a patent (UGARF case #1095) using this enzyme system in Azotobacter vinelandii. The chemical 1,3-propanediol is then used in place of ethylene in the production of polyesters to produce a polymer with superior properties for the fiber industry (currently marketed as Sorona by Dupont). A significant problem using our system as well as the B12-dependent system currently used by Dupont is that 1,3-propanediol is structurally similar to the metabolic substrate, glycerol, and therefore also a good inhibitor of both enzyme systems. By computational analysis of the available structural information we can predict site-specific mutations that will result in an enzyme with a lower affinity for the final commercial product, 1,3-propanediol, while not impacting the overall activity of the system.